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Vlugt, R.A.A. van der; Berendsen, M.

doi:10.1023/a:1015644409484

Cited by 68 publications

(26 citation statements)

References 11 publications

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“…Different strains of PepMV have been identified. The biological and molecular differences first observed between the tomato and pepino isolates of PepMV suggested that the tomato isolate of PepMV was a different strain, and was named 'tomato strain' (Van der Vlugt and Berendsen 2002). Further studies demonstrated that PepMV tomato strain isolates from different locations (Europe, North America and Canada) showed clear differences in symptom aggressiveness and nucleotide sequences when compared to the original pepino isolate (hereafter referred to as Peruvian isolate, PE) but only minor differences among them (Verhoeven et al 2003).…”

Section: Introductionmentioning

confidence: 99%

Simultaneous detection and identification of Pepino mosaic virus (PepMV) isolates by multiplex one-step RT-PCR

et al. 2009

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A RT-PCR was developed for the simultaneous detection and identification of three groups of Pepino mosaic virus (PepMV): European/Peruvian, Chilean 1/US1 and Chilean 2/US2 groups, followed by a restriction analysis that allowed the separation of the European, Peruvian, Chilean 2 and US2 isolates (patent pending). The multiplex RT-PCR reaction was performed by a mix of six primers that amplified a part of the RNA-dependent RNA polymerase gene of PepMV plus an internal control. Amplifications resulted in a 980 bp, 703 bp or 549 bp PCR product for European/Peruvian, Chilean 1/US1 or Chilean 2/ US2 groups, respectively. For the identification of the isolates present within the European/Peruvian and Chilean 2/US2 groups, the amplified PCR fragments were directly digested with SacI enzyme. The multiplex RT-PCR method presented higher sensitivity to detect CH1/US1 isolates in field samples than the RFLP-PCR method described by Hanssen et al. (European Journal of Plant Pathology 121:131-146, 2008). The detection limit observed with the multiplex RT-PCR was equal to or 3,125 times higher when compared to single RT-PCR or ELISA-DAS and molecular hybridisation methods, respectively. The use of the multiplex RT-PCR method in routine analysis of field tomato samples allowed the detection of 36.2 and 33.4% more positives when compared to the serological and molecular hybridisation methods, respectively, and the identification of plants infected with one, two or three isolates of PepMV.

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Section: Introductionmentioning

confidence: 99%

Simultaneous detection and identification of Pepino mosaic virus (PepMV) isolates by multiplex one-step RT-PCR

et al. 2009

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“…Sequences showed equal % similarity to both. Expected size (kb) of PCR product as reported by 1 van der Vlugt & Berendsen [11], and 2 Chen et al [12]. …”

Section: Resultsmentioning

confidence: 97%

“… 1 Degenerate primers of van der Vlugt & Berendsen [11], and 2 Chen et al [12]; 3 generic primer that anneals with first strand cDNA synthesis primer, KSDT. N = (A, G, C or T), D = (G+A+T), Y = (T or C), R = (A or G).…”

Section: Resultsmentioning

confidence: 99%

“…To give the best chance of detecting as many different types of viruses as possible, the procedure was carried out at a low annealing temperature with degenerate primers shown previously to amplify successfully members of the potex-, poty- and carlaviral families [11,12]. The clear and consistent differences in the amplification profiles obtained from the six colour-broken flowers compared with the non-colour-broken flowers (Figure 4A, B, C) was striking as the 30 cycles used in the PCR was expected to enable detection of very small quantities of viral RNA.…”

Section: Discussionmentioning

confidence: 99%

“…The degenerate and virus-specific primers used in PCR to amplify members of the potyvirus, potexvirus and carlavirus genera are listed in Table 4. The composition and utility of the degenerate primers was previously described in [11,12]. The PCR primer KS, complimentary to the 18 bp at the 5' end of the KSDT sequence was used to amplify viral sequences when a suitable reverse viral primer was not known.…”

Section: Methodsmentioning

confidence: 99%

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Colour break in reverse bicolour daffodils is associated with the presence of Narcissus mosaic virus

Hunter

Fletcher

Davies

et al. 2011

Virol J

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BackgroundDaffodils (Narcissus pseudonarcissus) are one of the world's most popular ornamentals. They also provide a scientific model for studying the carotenoid pigments responsible for their yellow and orange flower colours. In reverse bicolour daffodils, the yellow flower trumpet fades to white with age. The flowers of this type of daffodil are particularly prone to colour break whereby, upon opening, the yellow colour of the perianth is observed to be 'broken' into patches of white. This colour break symptom is characteristic of potyviral infections in other ornamentals such as tulips whose colour break is due to alterations in the presence of anthocyanins. However, reverse bicolour flowers displaying colour break show no other virus-like symptoms such as leaf mottling or plant stunting, leading some to argue that the carotenoid-based colour breaking in reverse bicolour flowers may not be caused by virus infection.ResultsAlthough potyviruses have been reported to cause colour break in other flower species, enzyme-linked-immunoassays with an antibody specific to the potyviral family showed that potyviruses were not responsible for the occurrence of colour break in reverse bicolour daffodils. Colour break in this type of daffodil was clearly associated with the presence of large quantities of rod-shaped viral particles of lengths 502-580 nm in tepals. Sap from flowers displaying colour break caused red necrotic lesions on Gomphrena globosa, suggesting the presence of potexvirus. Red necrotic lesions were not observed in this indicator plant when sap from reverse bicolour flowers not showing colour break was used. The reverse transcriptase polymerase reactions using degenerate primers to carla-, potex- and poty-viruses linked viral RNA with colour break and sequencing of the amplified products indicated that the potexvirus Narcissisus mosaic virus was the predominant virus associated with the occurrence of the colour break.ConclusionsHigh viral counts were associated with the reverse bicolour daffodil flowers that were displaying colour break but otherwise showed no other symptoms of infection. Narcissus mosaic virus was the virus that was clearly linked to the carotenoid-based colour break.

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Immunocapture-PCR for Plant Virus Detection

Mulholland

2008

Plant Pathology

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Untitled

Cited by 68 publications

References 11 publications

Simultaneous detection and identification of Pepino mosaic virus (PepMV) isolates by multiplex one-step RT-PCR

Simultaneous detection and identification of Pepino mosaic virus (PepMV) isolates by multiplex one-step RT-PCR

Colour break in reverse bicolour daffodils is associated with the presence of Narcissus mosaic virus

Immunocapture-PCR for Plant Virus Detection

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